Supplementary Materials

Ratiometric Sensing of Hydrogen Peroxide utilizing Conformational Change in Fluorescent Boronic Acid Polymers

Kan Takeshima,a Kanako Mizuno,a Hitoshi Nakahashi,a Horoshi Aoki,b and Yasumasa Kanekiyo*a

a Department of Biotechnology and Environmental Chemistry, Kitami Institute of Technology,

165 Koen-cho, Kitami, Hokkaido 090-8507, Japan.

b National Institute of Advanced Industrial Science and Technology (AIST), 16-1 Onogawa, Tsukuba, Ibaraki 305-8569, Japan.

Figure S1 1H NMR spectra of polymer B-10 measured in DMSO-d6. Assignments are the same as

those in Figure S2.

Figure S2 1H NMR spectra of polymer B-11 measured in DMSO-d6. Assignments are as follows:

9.701 ppm: amide (-CONH-), 8.211 and 8.045 ppm: aromatic (pyrene), 7.825-7.167 ppm: aromatic

(boronic acid), 6.824 ppm: amide (-CONH2), 4.952 ppm: methylene in pyrene unit.

Figure S3 1H NMR spectra of polymer B-12 measured in DMSO-d6. Assignments are the same as

those in Figure S2.

Figure S4 1H NMR spectra of polymer B-13 measured in DMSO-d6. Assignments are the same as

those in Figure S2.

Figure S5 1H NMR spectra of polymer B-01 measured in DMSO-d6. Assignments are as follows: 8.5

– 8.0 ppm: aromatic (pyrene), 7.5 – 6.8 ppm: aromatic (boronic acid) and amide (-CONH2), 5.014

ppm: methylene in pyrene unit.

Table S1 Elemental analysis data

SampleElemental data / wt%

C H N C / N (obs.) C / N (calc.)

B-10 55.16 5.84 6.53 8.45 8.15

B-11 52.98 6.14 9.33 5.68 5.70

B-12 51.10 6.36 10.60 4.82 4.90

B-01 50.37 7.32 15.67 3.25 3.21

Table S2 Estimated compositions of monomer units in copolymers

SampleMolar ratio / %

1 2 3

B-10* 92.2 0 7.8

B-11* 46.9 48.3 4.8

B-12* 30.3 65.0 4.8

B-01* 0 96.6 4.4*Molar ratio of 3 is supposed to be the same as in the feed solutions

Figure S6 Fluorescence spectra of polymer B-10 (1 mg L-1) excited at 348 nm in an aqueous

solution buffered at pH 10.9 (10 mM CAPS) at various H2O2 concentrations.

Figure S7 Fluorescence spectra of polymer B-11 (1 mg L-1) excited at 348 nm in an aqueous solution

buffered at pH 9.3 (10 mM CHES) at various H2O2 concentrations.

Figure S8 Fluorescence spectra of polymer B-11 (1 mg L-1) excited at 348 nm in an aqueous solution

buffered at pH 9.7 (10 mM CHES) at various H2O2 concentrations.

Figure S9 Fluorescence spectra of polymer B-11 (1 mg L-1) excited at 348 nm in an aqueous solution

buffered at pH 10.1 (10 mM CAPS) at various H2O2 concentrations.

Figure S10 Fluorescence spectra of polymer B-11 (1 mg L-1) excited at 348 nm in an aqueous

solution buffered at pH 10.5 (10 mM CAPS) at various H2O2 concentrations.

Figure S11 Fluorescence spectra of polymer B-11 (1 mg L-1) excited at 348 nm in an aqueous

solution buffered at pH 10.9 (10 mM CAPS) at various H2O2 concentrations.

Figure S12 Fluorescence spectra of polymer B-11 (1 mg L-1) excited at 348 nm in an aqueous

solution buffered at pH 11.3 (10 mM CAPS) at various H2O2 concentrations.

Figure S13 Fluorescence spectra of polymer B-12 (1 mg L-1) excited at 348 nm in an aqueous

solution buffered at pH 10.9 (10 mM CAPS) at various H2O2 concentrations.

Figure S14 Fluorescence spectra of polymer B-13 (1 mg L-1) excited at 348 nm in an aqueous

solution buffered at pH 10.9 (10 mM CAPS) at various H2O2 concentrations.

Figure S15 Fluorescence spectra of polymer B-01 (1 mg L-1) excited at 348 nm in an aqueous

solution buffered at pH 10.9 (10 mM CAPS) at various H2O2 concentrations.

Figure S16 Fluorescence excitation spectra of polymer B-11 (1 mg L-1) observed at 397 nm and 486

nm in an aqueous solution buffered at pH 10.9 (10 mM CAPS) in the absence of H2O2. The spectra

are normalized at 348 nm.

Figure S17 Absorption spectra of polymer B-11 (1 mg L-1) in an aqueous solution buffered at pH

10.9 (10 mM CAPS) at various H2O2 concentrations; inset shows the relationship between

absorbance at 290 nm and H2O2 concentration.

Figure S18 Time-course of excimer to monomer emission intensity ratio for an aqueous solution of

polymer B-11 (1 mg L-1) buffered at pH 10.9 (10 mM CAPS) containing 30 M H2O2.

Figure S19 Ratio of excimer to monomer emission intensities as a function of H2O2 concentrations

for aqueous solutions of polymer B-11 (1 mg L-1) buffered at pH 10.9 (10 mM CAPS) at various

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X : parts per Million : Proton8.0 7.9 7.8 7.7 7.6 7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1 6.0

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X : parts per Million : Proton8.0 7.9 7.8 7.7 7.6 7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1 6.0

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X : parts per Million : Proton8.0 7.9 7.8 7.7 7.6 7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1 6.0

(tho

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X : parts per Million : Proton8.0 7.9 7.8 7.7 7.6 7.5 7.4 7.3 7.2 7.1 7.0 6.9 6.8 6.7 6.6 6.5 6.4 6.3 6.2 6.1 6.0

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Field_Strength = 14.09636928[T] (600[MHz])X_Acq_Duration = 1.4548992[s]X_Domain = 1HX_Freq = 600.1723046[MHz]X_Offset = 5[ppm]X_Points = 16384X_Prescans = 1X_Resolution = 0.68733284[Hz]X_Sweep = 11.26126126[kHz]X_Sweep_Clipped = 9.00900901[kHz]Irr_Domain = ProtonIrr_Freq = 600.1723046[MHz]Irr_Offset = 5[ppm]Tri_Domain = ProtonTri_Freq = 600.1723046[MHz]Tri_Offset = 5[ppm]Clipped = FALSEScans = 8Total_Scans = 8

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---- PROCESSING PARAMETERS ----dc_balance( 0, FALSE )sexp( 0.2[Hz], 0.0[s] )trapezoid( 0[%], 0[%], 80[%], 100[%] )zerofill( 1 )fft( 1, TRUE, TRUE )machinephaseppmreference( -0.17757[ppm], 0[ppm] )

OH

B(OH)2

ppm

ppm

a

b

concentrations of NaCl.

Figure S20 1H NMR spectra of (a) 5 mM phenylboronic acid, and (b) 5 mM phenylboronic acid +

10 mM H2O2 measured in D2O containing 50 mM Na2CO3.

Figure S21 1H NMR spectra of (a) B-11, and (b) B-11 + 20 mM H2O2 measured in D2O containing

50 mM Na2CO3. Polymer concentration was 2 g L-1 ([boronic acid] = 7 mM).